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Introduction: Neonatal Septicemia is an important cause of morbidity and mortality. As infections due to ESBL producing K. pneumoniae & E. coli are on the rise, the present study was carried out in the NICU of KIMS, Narketpally, with an aim to identify any environmental sources & the mode of transmission over a period of 3 years from August 2006 to July 2009. Materials and Methods: A total of 264 neonates admitted with clinical features suggestive of septicemia in the NICU were studied by blood culture and CRP estimation. Antibiotic susceptibility pattern was determined. ESBL detection was done by double disc synergy test. Environmental samples from various sites (Incubators, phototherapy units, suction apparatus, trolley, door, floor, work surfaces) were collected using sterile swabs every month and processed simultaneously. Results: Of the 264 blood cultures, 197 (75%) showed bacterial growth. K. pneumoniae, 64 (32.7%) was the commonest organism followed by E. coli 55 (28%), S. aureus 31 (16%), Pseudomonas aeruginosa 28 (14%), Acinetobacter 13 (7%), and Coagulase negative Staphylococci 6 (2.8%) respectively. K. pneumoniae & E. coli were isolated from various environmental sites at least on one occasion and consistently from phototherapy units, door & floor of the NICU. The similarity between antibiograms of ESBL producing strains of K. pneumoniae & E. coli isolates from neonates and environment of NICU were statistically significant (P < 0.05). Conclusion: Wide spread use of third generation cephalosporins as a preemptive antibiotic for suspected cases of septicemia have contributed to the emergence of ESBL producing K. pneumoniae & E. coli in addition to other risk factors, both of which have extensively colonized the environment of the NICU. Repeated isolation of these two organisms from the NICU environment proves that some of the neonatal infections may be from the environment itself. Transmission can be stopped by maintaining the sterility of the NICU & hand hygiene among the mothers and health care workers.

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